Stool Total RNA Purification Kit – 49500

Stool Total RNA Purification Kit: Reliable, Inhibitor-Free RNA Extraction

The Stool Total RNA Purification Kit provides a rapid and robust method for the isolation of total RNA from human or animal stool samples. Extracting high-quality RNA from faecal matter is notoriously challenging due to the high concentration of PCR inhibitors, such as humic acids, bile salts, and complex polysaccharides. This kit overcomes these obstacles, delivering pure, ready-to-use RNA suitable for sensitive downstream applications without the need for toxic phenol-chloroform extractions.

Utilising a proprietary silicon carbide resin matrix, this kit employs an efficient spin-column chromatography method to bind and purify RNA while effectively removing contaminants. The process begins with rapid lysis and homogenisation of the stool sample, followed by the specific binding of RNA to the resin. Through a series of highly optimised wash steps, inhibitors and cellular debris are completely eliminated. The purified total RNA—encompassing both host and microbial profiles—is then eluted in a low-volume buffer, ensuring high concentration and maximum preservation of transcript integrity for subsequent gene expression analysis, microbiome studies, and pathogen detection.

Key Benefits:

• Complete Inhibitor Removal: Consistent downstream success. Optimised buffers and resin technology ensure the complete removal of humic acids and other faecal inhibitors, yielding RNA perfectly suited for sensitive RT-PCR, qPCR, and Next Generation Sequencing (NGS).
• Comprehensive RNA Profile: True representation of your sample. Extracts true total RNA, including both large mRNA and ribosomal RNA, as well as small RNAs (microRNA) from both the host and the microbiome without bias.
• High Yields and Purity: Maximum recovery from small inputs. Achieve superior yields of pure RNA from as little as 200 mg of stool, outperforming competitor kits in both quantity and optimal A260:A280/A260:A230 ratios.
• Phenol-Free Protocol: Safer laboratory environment. Eliminates the need for hazardous organic solvents like TRIzol or chloroform, providing a safer, more streamlined workflow.

Technical Insights

▼ View Technical Performance Data & Figures

Figure 1 Figure 1. High yields of stool total RNA were purified from 200 mg of human stool using Norgen’s Stool RNA Purification Kit and a leading competitor’s kit, in triplicate. For analysis, 7.5 µL of each 75 µL elution was loaded on a 1.2 % 1x MOPS formaldehyde-agarose gel. Norgen’s kit was found to have a higher yield of RNA, isolated from 200 mg of human infant stool.

Figure 2 Figure 2. Yield and Quality of Purified Stool RNA. Total RNA was isolated from 200 mg of human stool using Norgen’s Stool RNA Purification Kit and a leading competitor kit. Comparisons were then made based on yield, and A260:A280/A260:A230 ratios measured using the NanoVue Plus™. In Panel A it can be seen that both kits isolated RNA with high A260:A280 ratios (all samples were found to be above 1.8 and below 2.2). In Panel B, Norgen’s kit was found to isolate RNA with a high A260:A230 (with all samples once again falling in the 1.8-2.2 range). The competitor kit, however, was found to isolate RNA with extremely low A260:A230 ratios, with none of the samples displaying a A260:A230 ratio higher than 0.20. The results in Panel C are in agreement with the gel photo from Figure 1, and Norgen’s kit was found to isolate higher amounts of RNA, with an average yield of 14.58 µg, compared to the competitor’s average of 12.26 µg.

Figure 3 Figure 3. Detection of Bacterial Stool RNA using 16S Primers. Total stool RNA was isolated from 200 mg human stool samples using Norgen’s Stool RNA Purification Kit and a leading competitor’s kit. Five microliters of purified RNA was used in a 20 µL reverse-transcription reaction using Invitrogen’s Superscript III system with 16S reverse primers. The cDNA generated was then used in a qPCR reaction involving Norgen’s 2X PCR Mastermix spiked with SYBR green (Bio-Rad), using 0.3µM of primers against bacterial 16S. As can be seen in the amplification plot, Norgen’s kit outperformed the leading competitor’s kit by on average 1.5 Ct values. This indicates that Norgen isolated higher quality and yields of RNA from stool, that can be used in an array of downstream applications.

Specifications

• Maximum Sample Input: 200 mg of faecal matter
• Time to Complete: ~30 minutes
• Elution Volume: 50 µL
• Format: Spin Column
• Average Yield: ~14.5 µg (varies based on sample composition)
• Storage Conditions: Room temperature (15-25°C)